SMART SENTINEL: Nondestructive Point-of-Need CRISPR-Cas12a Lateral Flow Platform for Rapid New World Screwworm Field Triage
- 1Department of Chemistry and Biochemistry
- 2Texas Tech University
- 3School of Veterinary Medicine
Abstract
The recent re-emergence of the New World Screwworm (NWS) threatens the multibillion-dollar livestock industry, but current diagnostics require centralized morphological confirmation, creating severe 3 7-day testing bottlenecks due to an influx of non-target blowflies. To address this gap, we developed a rapid point-of-need (PON) CRISPR-Cas12a diagnostic framework for immediate NWS field triage. This assay features a nondestructive DNA extraction protocol that effectively isolates the target NWS cox1 gene while strictly preserving morphological integrity for mandatory federal verification. The assay uses the collateral cleavage activity of Cas12a to highly specifically cleave DNA probes upon Cas12a binding to the target cox1 gene via guide RNA. By combining a hand-held thermal cycler for target enrichment with lateral flow assay (LFA) readouts and portable digital imaging hardware, the integrated system couples molecular recognition with automated analysis to eliminate subjective field interpretation. To ensure biosecurity during development, we safely validated the pipeline using the Secondary Screwworm (C. macellaria) as a reliable congeneric surrogate proxy. Our assay demonstrates exceptional analytical sensitivity, establishing a limit of detection of 0.83 aM for synthetic targets, and exhibits absolute specificity against common distractor fly species, including Secondary Screwworms, Blue Bottle flies, Green Bottle flies, and House flies. Validation within a complex biological tissue matrix using a simulated beef liver infestation model confirmed robust diagnostic performance. Ultimately, this decentralized platform rapidly screens out negative samples, preventing the flooding of central reference laboratories and providing a critical biosecurity tool for swift epidemiological containment.
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BibTeX
@misc{garcia2026smart,
title = {{SMART SENTINEL: Nondestructive Point-of-Need CRISPR-Cas12a Lateral Flow Platform for Rapid New World Screwworm Field Triage}},
author = {Brianna Garcia and Jeshua Podliska and Zhicheng Peng and Minhao Liu and Isabela Bejgel de Araujo and Chad L. Cross and Olufemi Akinkuotu and Hui Chen},
howpublished = {ChemRxiv},
year = {2026},
month = aug,
doi = {10.26434/chemrxiv.15008044/v1},
url = {https://chemrxiv.org/doi/full/10.26434/chemrxiv.15008044/v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - SMART SENTINEL: Nondestructive Point-of-Need CRISPR-Cas12a Lateral Flow Platform for Rapid New World Screwworm Field Triage
AU - Brianna Garcia
AU - Jeshua Podliska
AU - Zhicheng Peng
AU - Minhao Liu
AU - Isabela Bejgel de Araujo
AU - Chad L. Cross
AU - Olufemi Akinkuotu
AU - Hui Chen
PY - 2026
DA - 2026/08/30
PB - ChemRxiv
DO - 10.26434/chemrxiv.15008044/v1
UR - https://chemrxiv.org/doi/full/10.26434/chemrxiv.15008044/v1
AB - The recent re-emergence of the New World Screwworm (NWS) threatens the multibillion-dollar livestock industry, but current diagnostics require centralized morphological confirmation, creating severe 3 7-day testing bottlenecks due to an influx of non-target blowflies. To address this gap, we developed a rapid point-of-need (PON) CRISPR-Cas12a diagnostic framework for immediate NWS field triage. This assay features a nondestructive DNA extraction protocol that effectively isolates the target NWS cox1 gene while strictly preserving morphological integrity for mandatory federal verification. The assay uses the collateral cleavage activity of Cas12a to highly specifically cleave DNA probes upon Cas12a binding to the target cox1 gene via guide RNA. By combining a hand-held thermal cycler for target enrichment with lateral flow assay (LFA) readouts and portable digital imaging hardware, the integrated system couples molecular recognition with automated analysis to eliminate subjective field interpretation. To ensure biosecurity during development, we safely validated the pipeline using the Secondary Screwworm (C. macellaria) as a reliable congeneric surrogate proxy. Our assay demonstrates exceptional analytical sensitivity, establishing a limit of detection of 0.83 aM for synthetic targets, and exhibits absolute specificity against common distractor fly species, including Secondary Screwworms, Blue Bottle flies, Green Bottle flies, and House flies. Validation within a complex biological tissue matrix using a simulated beef liver infestation model confirmed robust diagnostic performance. Ultimately, this decentralized platform rapidly screens out negative samples, preventing the flooding of central reference laboratories and providing a critical biosecurity tool for swift epidemiological containment.
ER -